Ginsenoside-Rg1 combined with a conditioned medium from induced neuron-like hUCMSCs alleviated the apoptosis in a cell model of ALS through regulating the NF-κB/Bcl-2 pathway / 中国天然药物

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease affecting both upper and lower motor neurons in the brain and spinal cord. One important aspect of ALS pathogenesis is superoxide dismutase 1 (SOD1) mutant-mediated mitochondrial toxicity, leading to apoptosis in neurons. This study aimed to evaluate the neural protective synergistic effects of ginsenosides Rg1 (G-Rg1) and conditioned medium (CM) on a mutational SOD1 cell model, and to explore the underlying mechanisms. We found that the contents of nerve growth factor, glial cell line-derived neurotrophic factor, and brain-derived neurotrophic factor significantly increased in CM after human umbilical cord mesenchymal stem cells (hUCMSCs) were exposed to neuron differentiation reagents for seven days. CM or G-Rg1 decreased the apoptotic rate of SOD1G93A-NSC34 cells to a certain extent, but their combination brought about the least apoptosis, compared with CM or G-Rg1 alone. Further research showed that the anti-apoptotic protein Bcl-2 was upregulated in all the treatment groups. Proteins associated with mitochondrial apoptotic pathways, such as Bax, caspase 9 (Cas-9), and cytochrome c (Cyt c), were downregulated. Furthermore, CM or G-Rg1 also inhibited the activation of the nuclear factor-kappa B (NF-κB) signaling pathway by reducing the phosphorylation of p65 and IκBα. CM/G-Rg1 or their combination also reduced the apoptotic rate induced by betulinic acid (BetA), an agonist of the NF-κB signaling pathway. In summary, the combination of CM and G-Rg1 effectively reduced the apoptosis of SOD1G93A-NSC34 cells through suppressing the NF-κB/Bcl-2 signaling pathway (Fig. 1 is a graphical representation of the abstract).

Metallurgic behavior and mechanical property of nickel-titanium endodontic files made by 3 heat treatment techniques / 中华口腔医学杂志

Objective@#To compare the phase transformation behavior, bending property and cyclic fatigue resistance of CM-wire, R-phase and M-wire nickel-titanium endodontic files treated by 3 heat treatment techniques.@*Methods@#Hyflex CM (25 mm, 25#/0.06) made from CM wire, TF (25 mm, 25#/0.06) made from R-phase heat treatment, ProTaper Next X2 (25 mm, tip size 25#/0.06) made from M-wire, Mtwo (25 mm, 25#/0.06) and ProTaper F2 (25 mm, tip size 25#/0.08) made from conventional nickel-titanium wire were chosen. Five of each files were chosen for differential scanning calorimetric (DSC) evaluation (n=5). The specimens were prepared from adjacent portions of the shaft. The weight and length were typically (20±1) mg and 2-3 mm. Another eight of the 5 nickel-titanium files were undergone cantilever-bending test, respectively (n=8). Further ten of the 5 instruments were submitted to the cyclic fatigue test by using a simulated metal root canal (n=10). The data of Ms, Mf, As, Af, ΔH, bending load, number of cyclic fatigue (NCF), fragment length were analyzed using one-way analysis of variance at a significance level of P<0.05.@*Results@#The Af of Hyflex CM [(60.27±0.94) ℃] and ProTaper Next [(51.40±0.32) ℃] were higher than body temperature of 37 ℃. The bending load values at 3.0 mm deflection of Hyflex CM [(0.867±0.074) N], TF [(1.275±0.146) N], Mtwo [(2.281±0.426) N], ProTaper Next [(3.104±0.252) N] and ProTaper [(4.227±0.483) N] increased gradually and significantly. The NCF of Hyflex CM (582±99), TF (427±85), ProTaper Next (158±22), Mtwo (129±32) and ProTaper (65±20) decreased gradually.@*Conclusions@#The 3 heat treatment techniques exert tremendous influence on the phase transformation behavior and mechanical characteristics of the files, which demonstrate improved bending property and cyclic fatigue resistance than the conventional ones.

Follistatin-like protein 1 plays a tumor suppressor role in clear-cell renal cell carcinoma / 癌症

BACKGROUND@#We previously showed that the expression of follistatin-like protein 1 (FSTL1) was significantly down-regulated in metastatic clear-cell renal cell carcinoma (ccRCC). In this study, we aimed to characterize the role of FSTL1 in the development of ccRCC.@*METHODS@#The effects of FSTL1 on cell activity and cell cycle were investigated in ccRCC cell lines with altered FSTL1 expression. Gene expression microarray assays were performed to identify the major signaling pathways affected by FSTL1 knockdown. The expression of FSTL1 in ccRCC and its effect on postoperative prognosis were estimated in a cohort with 89 patients.@*RESULTS@#FSTL1 knockdown promoted anchorage-independent growth, migration, invasion, and cell cycle of ccRCC cell lines, whereas FSTL1 overexpression attenuated cell migration. FSTL1 knockdown up-regulated nuclear factor-κB (NF-κB) and hypoxia-inducible factor (HIF) signaling pathways, increased epithelial-to-mesenchymal transition, up-regulated interleukin-6 expression, and promoted tumor necrosis factor-α-induced degradation of NF-κB inhibitor (IκBα) in ccRCC cell lines. FSTL1 immunostaining was selectively positive in epithelial cytoplasm in the loop of Henle, and positive rate of FSTL1 was significantly lower in ccRCC tissues than in adjacent renal tissues (P < 0.001). The multivariate Cox regression analysis showed that the intratumoral FSTL1 expression conferred a favorable independent prognosis with a hazard ratio of 0.325 (95% confidence interval 0.118-0.894). HIF-2α expression was negatively correlated with FSTL1 expression in ccRCC specimens (r = - 0.229, P = 0.044). Intratumoral expression of HIF-2α, rather than HIF-1α, significantly predicted an unfavorable prognosis in ccRCC (log-rank, P = 0.038).@*CONCLUSIONS@#FSTL1 plays a tumor suppression role possibly via repressing the NF-κB and HIF-2α signaling pathways. To increase FSTL1 expression might be a candidate therapeutic strategy for metastatic ccRCC.

Early growth response gene-1 regulates host cell autophagy in HTLV-1 infection / 中华微生物学和免疫学杂志

Objective To investigate the influence of early growth response gene-1 (EGR1) on the autophagy of host cells following infection with human T cell leukemia virus type 1 (HTLV-1).MethodsA HTLV-1-positive cell line MT2 was co-cultured with HeLa cells for 24 h to construct the virus early infection model.Immunoblotting assay was used to detect the expression of HTLV-1 core protein p19 and EGR1.Luciferase reporter gene analysis was used to detect the transcriptional activity of 5′-regulatory sequence of EGR1 at different time points after co-culturing.An effective small interfering RNA (siRNA) targeting EGR1 was screened out and transfected into HeLa cells by Lipofectamine 2000.Then the transfected HeLa cells were co-cultured with the HTLV-1-positive cell line MT2 for 24 h.Immunoblotting assay was used to detect HTLV-1 core protein p19, EGR1 and autophagy-related protein LC3.Real-time PCR was performed to detect viral load.Autophagosome was analyzed by immunofluorescence after co-culturing.Results The expression of EGR1 and the transcriptional activity of pEGR1-luc gradually increased after co-culturing HeLa cells with MT2 cells for 8 h (P<0.01).The expression of EGR1 was positively correlated with host cell autophagy following HTLV-1 infection.The effective siRNA for silencing the expression of EGR1 was obtained and named as siE2.The viral load, the expression of HTLV-1 core protein p19 and the proportion of LC3B/LC3A in the co-culture model were markedly down-regulated by RNA interference with siE2, which was concomitant with a persistent decrease of intracellular autophagosome (P<0.01).Conclusion EGR1 is associated with host cell autophagy and viral replication in HTLV-1 infection.

Effects of different techniques on removal of vapor lock in the apical region of curved canals: a cone-beam computed tomography study / 北京大学学报(医学版)

Objective:To compare the effect of four different techniques on removal of vapor lock in the apical region of curved root canals.Methods:Forty simulated resin root canals with 45° curvature were prepared using WaveOne Primary,then the apical foramen were sealed with soft wax.The teeth were divided randomly into 4 groups thereafter (n =10).Contract solution was injected into the canals using a 30 G side-vented needle and scanned with cone-beam CT (CBCT) to identify the volume of the vapor lock.Four different techniques including photon-induced photoacoustic streaming (PIPS) laser-activated irrigation,gutta-percha cone technique,ultrasonic irrigation,and sonic irrigation were used to remove the vapor locks in the root canals.The residual volume of the vapor lock was identified again using CBCT scanning data.Accordingly,the reduction rates of the vapor lock were calculated.Furthermore,the initial and residual vapor lock length was calculated.The data were analyzed by using the One-Way ANOVA analysis and Kruskal-Wallis H test at a significance level of P ＜ 0.05.Results:There was no significant difference in the initial vapor lock volume (P ＞ 0.05).Residual volume of the vapor lock for PIPS laser-activated irrigation was 0 mm3,and that for gutta-percha cone technique was (0.02 ± 0.07) mm3,significantly lower than those of ultrasonic and sonic irrigation,the values being (0.20 ± 0.09)mm3 and (0.23 ±0.06) mm3 (P ＜0.001),respectively.The reduction rates of the vapor lock of PIPS laser-activated irrigation and gutta-percha cone technique were 100.00％ (100.00％,100.00％) and 100.00％ (77.66％,100.00％),respectively,significantly higher than those of ultrasonic irrigation [70.37％ (56.41％,91.43％)] and sonic irrigation [63.54％ (51.47％,74.00％),P ＜0.001].The length of the residual vapor lock for PIPS laser-activated irrigation was 0 mm,and that for gutta-percha cone technique was (0.15 ±0.47) mm,significantly lower than those of ultrasonic and sonic irrigation,values being (2.21 ±0.09) mm and (2.34 ±0.08) mm (P ＜0.001),respectively.The length of the residual vapor locks in the ultrasonic and sonic group remained approximately the same as the distance between the working tip and the apical foramen.Conclusion:PIPS laser activated irrigation and gutta-percha cone technique could remove the vapor lock from the apical region of curved canals effectivelv.

Effect of the morphology of S-shaped root canals with reciprocating single file system on root canal irrigation / 中南大学学报(医学版)

Objective: To evaluate effect of the morphology of simulated S-shaped root canals with Reciproc or Mtwo instruments on root canal irrigation. Methods: A total of 40 simulated S-shaped resin blocks were randomly divided into 2 groups (n=20), which was prepared by Reciproc or Mtwo. Blue ink was injected with constant speed and volume into the canals using 30G lateral opening syringe. hTe distance from needle tip to apex and vapor lock length were measured. A type of 25# 0.04 gutta-percha point was used to stir and remove the vapor lock. hTe time for removal of the vapor lock was recorded. Another syringe was used to inject saline into the canals and to measure the length of stagnant water. Results: hTe distance from needle tip to apex in the Reciproc group was signiifcantly longer than that in the Mtwo group [(4.276 ± 0.221) mm vs (3.459 ± 0.205) mm,P<0.05]. The vapor lock length in the Reciproc group was signiifcantly longer than that in the Mtwo group [(4.472 ± 0.230) mm vs (3.668±0.217) mm,P<0.05]. hTe time to remove vapor lock was signiifcantly shorter in the Reciproc group than that in the Mtwo group [(10.13 ± 1.79) s vs (15.29 ± 2.15) s,P<0.05]. The length of stagnant water was also shorter in the Reciproc group than that in the Mtwo group[(1.351 ± 0.142) mm vs (2.245 ± 0.206) mm,P<0.05]. Conclusion: hTe morphology of S-shaped root canal affects the root canal irrigation. hTe effect of root canal irrigated by Reciproc is better than that by Mtwo.

Comparison of the shaping ability of reciprocating instruments in simulated S-shaped canals / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the shaping ability of Reciproc, WaveOne,Mtwo and ProTaper instruments in simulated S-shaped root canals.</p><p><b>METHODS</b>A total of 40 simulated S-shaped resin blocks were divided randomly into four groups, each group was prepared with Reciproc (group A), WaveOne (group B), Mtwo (group C) and ProTaper (group D), respectively. The preparation time and reduction of working length after preparation were measured. Pre- and postoperative images were obtained by a scanner and superimposed using Photoshop. Changes of coronal curve and apical curve curvature, as well as material removal from the inner and outer canal wall at 10 points beginning 1 mm from the end point of the canal, were measured using ImageJ. Centering ability was determined, accordingly. The data were analyzed using the one-way ANOVA and Student-Newman-Keuls.</p><p><b>RESULTS</b>The preparation time of group A and group B were (42.1 ± 2.7) and (41.5 ± 3.2) s respectively, significantly less than that of group C and group D [(62.7 ± 2.8), (62.8 ± 5.2) s] (P < 0.05).Reductions of working length after preparation were not significantly different among the four groups (P > 0.05). Coronal curve curvature changes of group A, group B and group D were (4.69 ± 0.63)°, (4.15 ± 0.89)° and (4.13 ± 0.59)° respectively, significantly less than that of group C[(5.26 ± 0.70)°], P < 0.05. Apical curve curvature changes were not significantly different among the four groups (P > 0.05). At the 2 mm point, the centering ability of group A[ (-0.242 ± 0.042) mm], group B[(-0.191 ± 0.077) mm], and group D[(-0.272 ± 0.046) mm] was better than group C[(-0.343 ± 0.057 mm] significantly (P < 0.05). At the 3 mm and 4 mm point, the centering ability of group A and group B was better than group C and group D significantly (P < 0.05). Whilst at the 5 mm point, the centering ability of group D was better than group A and group B (P < 0.05).</p><p><b>CONCLUSIONS</b>Reciproc and WaveOne could complete preparation faster and could maintain the original S-shaped canal curvature better than Mtwo and ProTaper, especially in the apical part.</p>

Comparison of the shaping capability of reciprocating instruments in simulated canals in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aims to evaluate the shaping capability of Reciproc, WaveOne, Mtwo, and ProTaper instruments in simulated root canals.</p><p><b>METHODS</b>A total of 40 simulated resin blocks were divided randomly into four groups. Each group was prepared with Reciproc (Group 1), WaveOne (Group 2), Mtwo (Group 3), and ProTaper (Group 4). The preparation time and reduction in working length after preparation were measured. Pre- and post-operative images were obtained with a scanner and superimposed through Photoshop. The changes in canal curvature and material removal from the inner and outer canal walls at 10 points beginning at 1 mm from the end point of the canal were measured with Image J. Centering capability was determined accordingly. Data were analyzed through one-way ANOVA, SNK, and Kruskal-Wallis at a significance level of P < 0.05.</p><p><b>RESULTS</b>The preparation time of Group 2 was (53.7 ± 6.7) s, whereas those of Groups 1, 3, and 4 were (86.9 ± 8.1) s, (112.2 ± 8.2) s, and (177.9 ± 11.2) s, respectively; the difference was found to be significant (P < 0.05). The reductions in working length among the four groups after preparation were not significantly different (P > 0.05). The canal curvature for Groups 1 to 4 were 2.671° ± 0.637°, 2.667° ± 0.450°, 3.664° ± 0.870°, and 3.797° ± 0.601°, respectively. The changes for Groups 1 and 2 were significantly smaller than those for Groups 3 and 4. At the 3 mm point, the transportation of Group 1 was (-0.016 ± 0.094) mm, which was significantly less than that of the other instruments (P<0.05). At the 4 mm and 5 mm points, the trans- portation values of Group 2 were (-0.080 ± 0.104) mm and (-0.312 ± 0.088) mm, which were significantly less than that of Group 1 [(-0.243 ± 0.099) mm, (-0.404 ± 0.064) mm, P < 0.05].</p><p><b>CONCLUSION</b>Reciproc and WaveOne can complete preparation faster and can maintain the original canal curvature better than Mtwo and ProTaper. Reciproc exhibits superior centering capability in the apical part of the canal, whereas WaveOne exhibits superior centering capability in the middle part of the canal.</p>

Prevalence and clinical characteristics of mitochondrial mutations in Chinese patients with type 2 diabetes / 中国糖尿病杂志

Objective To assess the prevalence of the A to G mutation at position 3243 of the mitochondrial tRNALeu(UUR) gene in type 2 diabetes in a Chinese population. Methods We screened 716 randomly selected, unrelated patients with type 2 diabetes for the mutation with a PCR-RFLP technique. Results Three individuals with this mutation were identified, representing approximately 0. 4％ of the type 2 patients screened. Further screening of first degree relatives of these 3 patients identified another 4 affected carriers. In comparison with type 2 diabetic patients without the mutation, these 7 carriers of the mt3243 mutation had:①an earlier onset of diabetes (38. 0±10.1 yr vs 53. 4±10.0 yr, P ＜0. 001) ;②lower BMI (19.5±2.0 vs 24. 9±10. 9, P ＜0. 0001) ;and ③ lower post-challenge insulin levels (Area under the curve of insulin levels during the OGTT, 2946± 1647.2 vs 7469±6647.7, P ＜ 0. 01). In addition, we screened the same 716 patients with type 2 diabetes, as well as 181 controls with normal glucose tolerance,for a newly described mt 3316 G→A mutation. This mutation was found in 16 patients with type 2 diabetes (2.20％) and 5 controls (2.7％). Therefore, the frequency of the mutation was not different between patients and controls.Moreover, clinical characteristics such as age of onset of diabetes, BMI, and insulin levels were not different between diabetic patients with the mt3316 mutation and those without it. Concision The mt3316 G→ A mutation is a polymorphism unrelated to diabetes.